KR100979888B1 - A composition comprising the powdered form or extract of ascidian Halocynthia roretzi as an active ingredient showing anti-diabetic activity - Google Patents

Abstract

The present invention is locust ( Halocynthia roretzi ) relates to a composition for the prevention and treatment of diabetes mellitus comprising the dry powder or extract of the extract as an active ingredient, specifically, the dry powder or extract of the larvae of the present invention of the animal model of diabetes induced by streptozotocin (STZ) Since the anti-diabetic effect is exhibited by significantly reducing blood sugar, the composition may be usefully used as a pharmaceutical composition or health functional food for preventing and treating diabetes.

Sludge, diabetes mellitus, streptozotocin

Description

A composition comprising the powdered form or extract of ascidian (Halocynthia roretzi) as an active ingredient showing anti-diabetic activity}

The present invention relates to a pharmaceutical composition or health functional food for the prevention and treatment of diabetes mellitus containing the dry powder or extract of the larvae as an active ingredient.

[1] Kang Jin-kyung, Concept of lifestyle-related diseases, Journal of Korean Medical Association, March. pp188-190, 2004

[Document 2] Health Insurance Review and Assessment Service, Health Insurance Review Statistics Index, 2003

[Reference 3] Ministry of Health and Welfare, National Health and Nutrition Survey, pp49-58, 2005

R. Brentjens et al., Surg . Clin North Am . 81 (3), pp 527-542, 2001

5 Z. Wang et al., Diabetes . 47 (1), pp 50-56, 1998

6 WJ Schnedl et al., Diabetes . 43 (11), pp 11326-33, 1994

7 A. Junod et al., J. Clin . Invert . 48, pp2129-2139, 1969

8 Grover JK et al., J. Ethnopharmacol . , 73, pp461-470, 2000

[Reference 9] 渡邊 勝 子, 鴻 巢 章 二.ホ ヤ の エ キ ス 成分.化學 と 生物. 27, pp 96, 1989

[Ref 10] Kang Ho-Ho et al., Journal of the Korean Fisheries Society, 26 (1), pp8-12, 1993

[Document 11] Kang Ho-Ho et al., Journal of the Korean Fisheries Society, 26 (2), pp150-158, 1993

[Reference 12] Lee Eung-Ho et al., Korean Journal of Food Science and Technology, 15 (1), pp1-5, 1983

13 K. Watanabe et al., Bull . Japan . Soc . Fish . 51, pp 1293, 1985

Document 14 CK Park et al., Nippon Suisan Gakkaishi . 56, pp 1319, 1990

15. K. Fujimato et al., Bull . Japan . Soc . Fish . 48, pp1323, 1982

16. BD Choi et al., J. Korean . Fish . Soc . 28, pp761, 1995

[Document 17] Hong, Byung-il et al., Journal of the Korean Fisheries Society. 35 (6), pp 671-675, 2002

[Document 18] Yuk Heung Sun et al., Korean Journal of Food and Nutrition. 32 (3), pp 474-478, 2003

19 Hsi-Hsien Liu et al., J. Agric . Food Chem . 50 (12), pp 3602-3606, 2002).

Document 20 i.水産食品成分の機能性について.月刊Food Chemical . 1989-2, pp 52-56, 1989

Document 21 AOAC. Offical Methods of Analysis. 16th ed. Association of Official Analytical Chemists, Washington DC., Pp69-74, 1995

[Reference 22] 日本 厚生 省.食品 衛生 指針 -I.日本 厚生 省 出版.東京. 30, 1960

26. E. Rabbo et al., Scandinav . J. Lab . Invest . 12, pp402-407, 1960

Modern man has lived a rich and convenient life with the development of industry. As a result, high levels of medicine have led to an increase in life expectancy, but on the other hand, a new disease called lifestyle-related disease has emerged. Lifestyle disease is a disease that has been renamed since 2003, which is commonly known as hypertension, allergies, obesity, diabetes, cirrhosis, heart disease, and cancer. The concept of lifestyle diseases was introduced because these diseases can be treated by improving eating habits, lifestyle, and environmental factors (Kang, Jin Kyung, Concept of lifestyle diseases, Korean Medical Association, March, pp. 188-19). 190, 2004).

Data on lifestyle-related diseases surveyed by the Health Insurance Review & Assessment Service showed that the number of patients using medical institutions increased from 2000 to osteoporosis (89.3%), high cholesterol (80.9%), high blood pressure (53.9%), diabetes (45.5%), and heart disease (29.0%). ). In particular, the average annual increase in the number of cases was high cholesterol (21.8%) and diabetes was 3.4% (Health Insurance Review and Assessment Service, Health Insurance Review and Statistics, 2003; Ministry of Health and Welfare, National Health and Nutrition Survey, pp49-58, 2005). Therefore, in Korea, measures for hyperlipidemia and diabetes-related diseases are urgently needed.

Diabetes mellitus is one of the most common chronic adult diseases, and it is estimated that about 5% of the total population in Korea is at least 2.5 million. In advanced countries, the number of diabetic patients is increasing rapidly each year. In Korea, the number of patients is expected to increase gradually as the standard of living and the lifestyle become western. Diabetes is a metabolic disorder in which the body's ability to use glycose normally impairs its blood sugar levels and increases the amount of glucose in the blood, causing excess sugar to be excreted in the urine. Diabetes is caused by a lack of insulin, and a lack of insulin occurs when the absolute amount of insulin is low, or when a relatively low amount of hormones other than insulin is released. Insulin, a polypeptide hormone, is produced from beta cells of the island of Langerhans in the pancreas. When pancreatic hormone insulin is balanced in the body with a hormone that acts against the insulin secreted by the pituitary gland, the adrenal gland and the thyroid gland, the glycemic control function is normal. maintain.

Diabetes mellitus is characterized by two types: insulin-dependent type 1 diabetes mellitus is caused by a deficiency in the secretion of insulin, a glycos regulatory hormone in the blood, mainly younger people in their 10s to 20s. It is also called juvenile diabetes because it occurs in. Type 2 diabetes mellitus occurs mainly after the age of 40 and accounts for most of the diabetic patients in Korea. Unlike type 1, it is called adult-type diabetes, and the cause of the disease is not clear yet, but it is known that it is caused by genetic factors and environmental factors. The etiology of type 2 diabetes is both a disorder of insulin secretion in pancreatic beta cells and a deficiency of insulin action (insulin resistance) in target cells, of which it is not yet clear which changes are of primary importance.

Streptozotocin (STZ) is a compound that inhibits the production of insulin in mammals and is used to induce type 1 diabetes in laboratory animals. The US FDA authorizes the use of streptozotocin for the treatment of pancreatic cancer, which can reduce the size of cancer cells or alleviate various symptoms caused by cancer when given to the patient (R. Brentjens et al., Surg Clin North Am . 81 (3), pp 527-542, 2001). However, there are problems due to the toxins of this substance, and even it causes cancer, so it is used on a limited basis. The amount used for inducing diabetes is usually injected 500 mg / m ² / day through intravenous injection for 5 days, and this is repeated for 4-6 weeks. The IUPAC name for streptozotocin is 1-methyl-1-nitroso-3- [2,4,5-trihydroxy-6- (hydroxymethyl) oxan-3-yl] -urea (1-methyl-1- nitroso-3- [2,4,5-trihydroxy-6- (hydroxymethyl) oxan-3-yl] -urea), a glucosamine-nitrosourea compound. Like other alkylating agents, nitrosourea compounds not only cause DNA damage, but also cause other harmful chemical reactions (Z. Wang et al., Diabetes . 47 (1), pp 50-56, 1998; WJ Schnedl et al., Diabetes . 43 (11), pp1326-33, 1994). One of them is the destruction of pancreatic beta-cells. It is reported that 7 hours after streptozotocin is administered, blood glucose is rapidly decreased and insulin concentration is increased, but insulin secretion is decreased due to the destruction of beta-cell, and blood sugar is gradually increased, and after 24 hours, hyperglycemic state is reported. (A. Junod et al., J. Clin . Invert . 48, pp2129-2139, 1969).

Agents currently used for the treatment of diabetes include five groups of compounds: biguanides, thiazolidinediones, sulfonylureas, benzoic acid derivative compounds and α-glucosidase. Inhibitors (α-glucosidase inhibitors) are being used, and a biguanide compound such as metformin is known to prevent excessive blood gluconeogenesis. Thiazolidinedione compounds are thought to act to increase glucose influx in peripheral tissues, benzoic acids such as sulfonylureas, repaglinide compounds such as tolbutamide and glyburide Derivatives stimulate insulin secretion, while α-glucosidase inhibitors such as acarbose act to lower carbohydrate glucose by inhibiting the digestion of carbohydrates in the small intestine.

However, since the preparations containing such compounds have side effects that cause nausea, vomiting, diarrhea and rash at the beginning of administration, an excellent hypoglycemic agent having low side effects and high safety is required. To date, there is no cure for diabetes, and there are many researches to search for diabetes treatment from natural products due to the side effects of drug treatment.Therefore, the 1990 World Health Organization (WHO) has no effect on diabetes. The use of natural and low side effects is highly recommended (Grover JK et al., J. Ethnopharmacol . , 73, pp461-470, 2000).

Sea urchins inhabit about 2,000 species worldwide, especially Halocynthia roretzi . ) Species is widely used in Korea and Japan for reproduction. In recent years, the production of squirrel is rapidly increasing due to the spread of squirrel farming technology, and many researches on wild squirrel are being conducted on wild and aquatic farms. (27, pp96, 1989 ). Studies on the food composition of the squirrel are as follows: Chemical composition of the squirrel (Lee Kang-Ho et al., 26 (1), pp8-12, 1993), 158, 1993; Lee Eung-ho et al., Korean Journal of Food Science and Technology, 15 (1), pp1-5, 1983), seasonal variation of nitrogen-containing and free amino acids (K. Watanabe et al., Bull . Japan.Soc . Fish . 51 , pp 1293, 1985; CK Park et al., Nippon Suisan Gakkaishi . 56, pp1319, 1990), and studies on precursors and production mechanisms of odorous odors (K. Fujimato et al., Bull . Japan . Soc . Fish . 48, pp1323, 1982), and Choi et al. Odor components have been analyzed and reported (BD Choi et al., J. Korean . Fish . Soc . 28, pp761, 1995).

As a report on the pharmacological effects of larvae, Hong et al. Reported that ACE (angiotensin converting enzyme) of polysaccharide sulfate-derived sulphate inhibits blood pressure and consequently decreases blood pressure. It has been reported that serum lipid content is lowered (Hong Byung-il et al., Journal of the Korean Fisheries Society. 35 (6), pp671-675, 2002; Yuk Heung-sun et al., Korean Journal of Food and Nutrition. Liu et al also has a reported sikindago the GAGs (glycosaminoglycans) distributed in many tissues, such as squirt promote the metabolism of lipid and improving the immune system (Hsi-Hsien Liu et al. , J. Agric. Food Chem . 50 (12), pp 3602-3606, 2002).

Of aquatic functionality it is very likely to act as a composite component than the component alone as in the herbal medicine (芦己勝朗.水産食品成分の機能性について. 月刊 Food Chemical . 1989-2, pp 52-56, 1989). Even if the assumption that functional properties will be expressed by such a complex component system is not correct, the separation process of functional food materials in aquatic products is mistaken for removing well-known functional materials. For example, in the case of chitin functional material, which is well known as a functional food material, in order to obtain only the final chitin component from the skin of the red crab, the minerals such as CaCO ₃ which prevents osteoporosis may be first removed through strong HCl pretreatment, and then Strong NaOH treatment removes astaxanthin, which has antioxidant capacity, and hydroxyl protein, a typical functional nutrient.

Therefore, the present inventors first select a squirrel to search for a material in aquatic products that makes anti-diabetic efficacy close to drugs as a complex component system, and examines whether it expresses a functional activity by itself without separating a specific functional ingredient so that the data can be compared. Under the same conditions, the present invention was completed by confirming the effect of reducing blood sugar through the animal experiment of diabetes induced by streptozotocin for functional verification.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus containing a dry powder or extract of the larvae as an active ingredient.

The present invention also provides a health functional food for the prevention and improvement of diabetes mellitus containing the dry powder or extract of the larvae as an active ingredient.

Halocynthia as defined herein roretzi) is dongmulmun notochord (Phylum Chordata), ascidians (Class Asidiacea ), Side Seaweed ( Order Pleurogona ) or Sea Urchin ( Family Pyuridae ) includes whole, meat or skin of the larvae that lives in Korea, Japan, etc.

Extracts as defined herein include solvents selected from water, including purified water, lower alcohols having 1 to 4 carbon atoms, or mixed solvents thereof, preferably water, ethanol, or mixed solvents thereof.

Diabetes as defined herein includes type 1 or type 2 diabetes, preferably type 1 diabetes.

Hereinafter, the present invention will be described in detail.

A first step of washing the squirrel of the present invention; A second step of separating and cutting the skin and the meat; A third step of quenching the same at -100 ° C to 0 ° C, preferably at -50 ° C to -30 ° C for about 6 hours to 24 hours, preferably about 12 hours; Through the manufacturing method comprising the fourth step of freezing under reduced pressure can be obtained the dried powder of the present invention in a dry state.

In addition, the extract of the larvae of the present invention can be prepared as follows. A first step of cutting the snails and freeze-drying them, including body fluids and intestines; This is a solvent selected from water containing purified water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, preferably water, ethanol or a mixed solvent thereof, at 0 ° C to 100 ° C, preferably at 20 ° C to 50 ° C. A second step of extracting for minutes to 1 hour, preferably for 30 minutes; This can be obtained through the production method comprising a third step of concentration under reduced pressure and freeze-dried.

The present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus containing the dry powder or extract of the larvae obtained by the above production method as an active ingredient.

In addition, the present invention provides a health functional food for the prevention and improvement of diabetes mellitus containing the dry powder or extract of the squirrel obtained by the above production method as an active ingredient.

The pharmaceutical composition for the prevention and treatment of diabetes mellitus containing the dry powder or extract of the larvae of the present invention comprises 0.1 to 50% by weight of the snail relative to the total weight of the composition.

The pharmaceutical composition containing the dry powder or extract of the larvae of the present invention may further include appropriate carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.

The pharmaceutical composition containing the snails of the present invention is formulated in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be used. Carriers, excipients and diluents which may be contained in the composition containing the compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose, etc. Or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of scavengers of the present invention may vary depending on the age, sex, and weight of the patient, but may be administered once to several times daily from 0.1 to 100 mg / kg. The dosage may also be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The pharmaceutical composition may be administered to various mammals such as mice, mice, livestock, humans, and the like. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a dietary supplement for the prevention and improvement of diabetes comprising a dry powder or extract of the snail as an active ingredient. Foods to which it may be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to foods or beverages for the purpose of preventing the prevention of diabetes. At this time, the amount of dry powder or extract of sea cucumber in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health functional beverage composition is 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be added.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the dry powder or extract as essential ingredients in the indicated ratios, and contains various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. can do. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tautin, stevia extract) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the dry powder or extract of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the dry powder or extract of the present invention may contain a pulp for the production of natural fruit juices and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the dry powder or extract of the present invention.

The present invention relates to a composition for the prevention and treatment of diabetes mellitus containing the dry powder or extract of the squirrel as an active ingredient. Specifically, the dry powder or extract of the squirrel of the invention is a blood sugar of a diabetic animal model induced by streptozotocin Significantly reduce the anti-diabetic effect by reducing the composition, the composition can be usefully used as a pharmaceutical composition or health functional food for the prevention and treatment of diabetes.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, but the content of the present invention is not limited thereto.

Example  1. Preparation of Sample

1-1. Sample Preparation

Locust ( Halocynthia roretzi ) was farmed off the coast of Yeongjin, Gangwon-do. On December 5, 2006, a total of 294 fish were purchased from Jumunjin Fish Market. The edible part and the shell were separated and used as a sample for the following experiment. The average weight of the larvae was 64.7 ± 10.2 g. Among them, the average weight of meat without shells was 50.5 ± 7.9 g, which was 78 ± 6.3%. The average diameter of the larvae was 48.4 ± 8.3 mm, and the average height was 106.1 ± 18.4 mm, and was used as a sample of the following experiment (hereinafter referred to as “HR”).

1-2. Preparation of Soluble Ethanol Soluble Extract

250 g of the squirts obtained in Example 1-1 were cut at 100 mm intervals, and the sample containing the bodily fluid and gut flowing out during the cutting process was freeze-dried under vacuum, and 1 L of 70% ethanol was added thereto for 30 minutes at 25 ° C. Extraction was performed once, concentrated with a vacuum concentrator, and lyophilized to obtain 8.2 g of ethanol soluble extract of squill (hereinafter referred to as "HR-1").

1-3. Manufacture of Dry Powder

250 g of the turmeric obtained in Example 1-1 was thoroughly washed with tap water to remove foreign substances and salt, and then supported by a sieve to minimize water, and the shell and meat were separated to be fed separately. . During the cutting process, the body fluids and intestines flow out, and the squid was included in the meat and spread flat and thin on the tray. At this time, about half the thickness of the sample in the tray, left in a fast freezing machine (DFS520) for 12 hours at -50 ℃ with a temperature sensor plugged in completely frozen, and then freeze-drying the pressure-reducing freeze dryer (Iljin, Korea Ilshinsa, PVIFD30A) ) The principle of the freeze drier is that when the tray is slowly heated, the sample is in a nearly vacuum state so that the phase changes from solid to gas. At this time, the water vapor of the sample is gradually reduced by collecting the water vapor by cooling it again. The conditions for the vacuum freeze dryer used in the experiment were vacuum up to 5 mili torr and temperature up to 30 ° C. In order to prevent the sample from bubbling (boiling and overflowing out of the tray) by rapid temperature rise, the temperature for heating the sample is gradually raised from 0 ° C. After about 42 hours, drying stops when the temperature of the sample and the tray reaches 30 ° C. To obtain a dry powder of the larvae (hereinafter referred to as "HR-2").

As a result of the experiment, as shown in Table 1, the HR obtained in Example 1-1 occupies 78.0% of the edible part, and 22.0% of the shell part of the non-edible part, whereas Example 1-3 The dry sample obtained in U.S. showed 42.8% of meat (hereinafter referred to as "HR-M") and 57.2% of shell part (hereinafter referred to as "HR-T"), resulting in a higher proportion of non-planted parts. Could confirm.

Weight of HR (%) Weight of HR-2 (%) Yield (%) HR-M 14,836 g (78.0%) 550 g (42.8%) 5.0 HR-T 4,185 g (22.0%) 734 g (57.2%) 13.1 all 19,021 g (100.0%) 1284 g (100.0%) 6.8

Reference Example  1. Experimental animals and Experimental  Ready

Experimental animals were distributed from Daehan Biolink Co., Ltd. (negative Chungbuk) 1 in the animal breeding room of the university under constant conditions (temperature: 22 ± 1 ℃, humidity: 55 ± 3%, contrast: 12 hours light / dark cycle) Sprague-Dawley male rats weighing 200 ± 10 g body weight adjusted to solid foods for 4 weeks were divided into 4 groups of 9 rats per group, the control group fed the normal diet, the negative control group induced by streptozotocin and the HR-2 administration. Classified as test group. HR-M and HR-T were suspended in 10% tween80, respectively, and 100 and 200 mg per kg of experimental animals were ingested once a day for 2 weeks and 4 weeks using an oral zone.

Experimental Example  1. General Ingredient Analysis

In order to analyze the general components of HR obtained in Example 1, experiments were performed using the AOAC method as follows (AOAC.Offical Methods of Analysis.16th ed.Association of Official Analytical Chemists, Washington DC., Pp69- 74, 1995).

Specifically, moisture was measured by atmospheric heating drying method, crude fat was measured by Soxthlet method, and ash was measured by dry calcination method. Crude protein was measured by semimicro Kjedahl method and nitrogen coefficient (6.25) was used. Calculated.

As a result, as shown in Table 2 and Table 3, HR had a very high content of moisture in both the edible and non-edible parts, and there were a lot of crude ash and polysaccharides in meat, and a lot of crude ash in non-edible portion. The above-mentioned HR-M contained almost equal amounts of polysaccharides, minerals, fats and proteins, and HR-T contained only a high content of 74% of minerals, followed by polysaccharides, proteins, and fats. In addition, HR-2 was composed mostly of minerals and proteins, contained polysaccharides, and had a very low crude fat content. HR showed a high moisture content of 84.5 to 93.4%.

ingredient HR-M HR-T moisture 90.7 ± 0.7 84.5 ± 0.8 Crude protein 1.8 ± 0.1 1.4 ± 0.03 Crude fat 2.4 ± 0.02 0.8 ± 0.02 View minutes 2.5 ± 0.4 11.5 ± 0.3 Polysaccharide 2.6 ± 0.01 2.0 ± 0.01

ingredient HR-M HR-T Crude protein 18.9 ± 1.1 8.7 ± 0.2 Crude fat 25.8 ± 2.2 4.8 ± 0.1 View minutes 27.3 ± 4.6 73.6 ± 1.6 Polysaccharide 28.1 ± 0.1 12.9 ± 0.1

Experimental Example 2. Determination of volatile basic nitrogen (VBN) content

In order to analyze the volatile basic nitrogen content of HR obtained in Example 1-1, a microdiffusion method using a Conway unit was carried out as follows. (日本 厚生 省. 食品 衛生 指針 -I. 日本 厚生 省出版. 東京. 30, 1960).

2 g of HR was added to the analysis beaker of volatile basic nitrogen, 16 ml of distilled water was added, and the mixture was stirred well with a glass rod. For protein precipitation, 2 mL of 20% TCA solution was added, ground well, and left to stand for 10 minutes to obtain a supernatant, which was then used as a sample solution. 1 mL of boric acid absorbent was added to the inner chamber of the Conway unit and 1 mL of the sample solution into the outer chamber. Then, the lid was closed for about 2/3. Then, 1 mL of saturated potassium carbonate solution was added to the outer chamber. The outer chamber liquid, that is, saturated potassium carbonate and the sample solution, were mixed while the unit was slowly rotated by inserting a clip so that the inner and outer chamber liquids did not mix with each other. Thereafter, the mixture was left at 37 ° C. for 80 minutes and titrated with 0.01 N HCl solution. Titrate until the hue of the solution changes from green to colorless to scarlet. In the black test, a 20% TCA solution was added to the outer chamber in the manner of the above conditions. 1 mL of 0.01 N HCl solution used corresponds to 0.14 mg of volatile basic nitrogen. The content of volatile basic nitrogen was calculated according to the following equation.

Volatile Basic Nitrogen  ( mg %) = 0.14 × (A-B) × 20 × f × 100 / S

   S: Sample Collection

   A: Proper amount of sample ( mL )

   B: Blank Proper amount of mL )

   f: 0.01 N HCl  Solution factor

VBN (Volatile basic nitrogen), that is, volatile basic nitrogen, is used to measure the freshness of fish and shellfish. In particular, it is used as a standard for the initial decay of aquatic products. 30 mg% of frozen fish, 50 mg% of sharks and skates, 20 mg% of pollack and cod, 20 mg% of shellfish and shellfish, and 30 mg% of other seafood.

As a result, the VBN value of HR obtained in Example 1-1 was 7.0 to 8.7 mg% level as shown in Table 4, all of which was found to be very fresh as it does not significantly fall below the initial decay criteria.

HR-M HR-T VBN (mg%) 8.7 ± 0.7 7.0 ± 0.2

Experimental Example 3 Diabetic Animal Experiment Induced by Streptozotocin (STZ)

To induce diabetes, 50 mg / kg of streptozotocin (STZ) was dissolved in 0.05 M citric acid buffer (pH 4.5) by the method of Wang, and then injected into the tail vein. Rats in the range of 300-400 mg / dV were measured and used as experimental animals in the following experiments (Z. Wang et al., Diabetes. 47 (1), pp 50-56, 1998).

3-1. How to measure blood sugar

Blood glucose was measured using a kit prepared according to the glucose oxidase method (Exactech) according to the method of Rabbo et al. (E. Rabbo et al., Scandinav. J. Lab. Invest . 12, pp402 -407, 1960).

3-2. Statistical processing

As shown in Table 5, the results obtained in this experiment were expressed as mean ± standard deviation, and a significant difference test was performed by applying Duncan's multiple range test, which shows the difference between each experimental group by PC-STAT program. It was. With this PC-STAT program, there can be a minimum of 2 to 50 significant difference test treatments and up to 150 experimental values. In other words, a maximum of 150 experimental values of several samples are inputted in one quotient, and two to 50 groups are compared to each other simultaneously to compare two sections to determine mutual significance.

As a result, as shown in Table 5, the HR-2 administration test group showed no significant difference from the negative control group at 2 weeks of administration, but the blood glucose level of the HR-2 administration test group was significantly higher than the negative control group at 4 weeks of administration. I could see that falling into. In particular, the HR-T-administered test group showed better blood glucose reduction effect than the HR-M-administered test group, and the HR-T 200 mg / kg-administered test group had a 23.9% decrease in blood glucose compared to the negative control at 4 weeks. Could confirm.

Test group Dose
(mg / kg)      Concentration (mg / dl) 2 weeks 4 weeks Control 93.5 ± 10.7 ^b 98.7 ± 9.57 ^d Negative Control (STZ) 380.2 ± 45.2 ^a 379.8 ± 25.4 ^a HR-M 100 365.6 ± 35.2 ^a 369.8 ± 30.1 ^ab 200 369.9 ± 27.8 ^a 368.2 ± 31.9 ^ab HR-T 100 381.7 ± 32.1 ^a 345.3 ± 20.5 ^ab 200 376.9 ± 25.4 ^a 289.1 ± 18.6 ^c Values marked with the same letter are not significantly different from each other (p <0.05, Duncan's multiple range test)

It describes a formulation example of a composition comprising a dry powder or an extract of the present invention, the present invention is not intended to limit it, but is intended to explain in detail only.

Formulation Example 1 Preparation of Powder

HR-M 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

HR-T 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are usually prepared by tableting according to the method for preparing tablets.

Formulation Example 3 Preparation of Capsule

HR-M 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

HR-T 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

It is usually prepared in the above ingredient content per ampoules (2 ml) according to the preparation method of the injection.

Formulation Example 5 Preparation of Liquid

HR-M 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified Water Proper Amount

Normally, each component is added to the purified water to dissolve according to the preparation method of the liquid solution, lemon flavor is added, the above ingredients are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled into a brown bottle and sterilized. To prepare a liquid solution.

HR-T 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinamide 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5 mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

15 mg potassium monophosphate

Dicalcium Phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is a composition that is relatively suitable for the health functional food, the composition is mixed in a preferred embodiment, but the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

HR-M 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

900 ml of purified water whole

After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored Used to prepare the healthy beverage composition of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (6)

A first step of rapidly quenching the shell of Korean barley for 6 to 24 hours at -100 to 0 ° C; A pharmaceutical composition for preventing and treating type 1 diabetes, comprising the dry powder obtained through the second step of the process of freezing this under reduced pressure as an active ingredient. delete delete delete delete delete